聚乙二醇修饰牛血红蛋白产物的分离纯化和检测

Separation,Purification and Detection of Product of Bovine Hemoglobin Modified by Polyethylene Glycol

目的:研究聚乙二醇修饰剂单甲氧基聚乙二醇琥珀酰亚胺酯(SC-mPEG)对牛血红蛋白(BHB)修饰产物的合适的分离纯化、检测方法。方法:采用阳离子交换层析以及有机溶剂沉淀法进行分离,并采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和三硝基苯磺酸(TNBS)法进行检测。结果:阳离子交换层析法分离出修饰与低修饰产物,TNBS法检测阳离子交换层析分离所得2个峰值产物的平均修饰率分别为52.95%和14.19%。结论:阳离子交换层析法可作为分离纯化SC-mPEG-BHB的方法;TNBS方法可作为检测SC-mPEG-BHB平均修饰率的方法。

OBJECTIVE： Researching the appropriate methods to separation, purification and detection the mixture of PEG modifier SC- mPEG with BHB. METHODS： Adopting cation- exchange chromatography and organic solvent precipita- tion to separate the mixture, then detecting it by SDS - PAGE and TNBS methods. RESULTS： Modifer and non - modifer product were separation by cation - exchange chromatography. The average modification rates of two peak product from cation- exchange chromatography are 52.95% and 14.19% detected by TNBS methods . CONCLUSIONS： Cation- exchange chromatography methods could be to separate and purify the SC - mPEG - BHB, TNBS methods coule be to detect the average modification rate of the SC-mPEG -BHB.